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Biosafety studies of carrier cells infected with a replication-competent adenovirus introduced by IAI.3B promoter.

Hamada K, Shirakawa T, Terao S, Gotoh A, Tani K, Huang W - Mol Ther Methods Clin Dev (2014)

Bottom Line: The use of carrier cells infected with oncolytic viruses in cancer gene therapy is an attractive method because it can overcome viral immunogenicity and induce tumor immunity and significant antitumor activity.Freeze-thawing increased the antitumor effect of A549 carrier cells by promoting the translocation of oncolytic adenovirus particles from the nucleus to the cytoplasm following the rupture of the nuclear membranes.In chronic toxicity tests in rabbits, there were no serious side effects after eight doses of 1.25 × 10(7) cells/kg or less for 4 weeks; a significant immune response is known to elicit increased numbers of antiadenovirus antibodies and enlarge the spleen.

View Article: PubMed Central - PubMed

Affiliation: Department of Obstetrics and Gynecology, School of Medicine, Ehime University , Shitsukawa, Toon, Ehime Japan.

ABSTRACT
The use of carrier cells infected with oncolytic viruses in cancer gene therapy is an attractive method because it can overcome viral immunogenicity and induce tumor immunity and significant antitumor activity. To enable human clinical trials of this treatment, acute and chronic toxicity tests must first be performed to ensure safety. IAI.3B promoter, oncolytic adenovirus AdE3-IAI.3B introduced by IAI.3B promoter, and A549 carrier cells infected with AdE3-IAI.3B were highly active in cancer cells but not in normal cells. Freeze-thawing increased the antitumor effect of A549 carrier cells by promoting the translocation of oncolytic adenovirus particles from the nucleus to the cytoplasm following the rupture of the nuclear membranes. No deaths or abnormal blood test data resulted from acute toxicity tests conducted in nude mice after a single dose. In chronic toxicity tests in rabbits, there were no serious side effects after eight doses of 1.25 × 10(7) cells/kg or less for 4 weeks; a significant immune response is known to elicit increased numbers of antiadenovirus antibodies and enlarge the spleen. From these results, it could be concluded that cancer gene therapy of recurrent solid tumors using carrier cells can be safely trialed in humans.

No MeSH data available.


Related in: MedlinePlus

Infection and storage condition of AdE3-IAI.3B-infected A549 carrier cells. (a) In vitro growth inhibitory effect of A549 carrier cells infected with AdE3-IAI.3B at 200 MOI and irradiated at 200 Gy. (b) Effects of radiation exposure on tumorigenicity of A549 cells subcutaneously transplanted into nude mice (n = 5 per group). (c) Effects of infection time, antiadenovirus antibodies and freeze-thawing on the cytotoxicity of AdE3-IAI.3B-infected A549 carrier cells in ovarian cancer HEY cells. *P < 0.05; **P < 0.01. (d) Changes in viability of AdE3-IAI.3B-infected A549 carrier cells by infection time. *P < 0.05. (e) Changes of PFU activity of AdE3-IAI.3B-infected A549 carrier cells by infection time. *P < 0.05. (f) Changes of viability of AdE3-IAI.3B-infected A549 carrier cells following liquid nitrogen storage for 3 months. (g) Changes of PFU activity in cells and supernatants of AdE3-IAI.3B-infected A549 carrier cells following liquid nitrogen storage for 3 months.
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fig2: Infection and storage condition of AdE3-IAI.3B-infected A549 carrier cells. (a) In vitro growth inhibitory effect of A549 carrier cells infected with AdE3-IAI.3B at 200 MOI and irradiated at 200 Gy. (b) Effects of radiation exposure on tumorigenicity of A549 cells subcutaneously transplanted into nude mice (n = 5 per group). (c) Effects of infection time, antiadenovirus antibodies and freeze-thawing on the cytotoxicity of AdE3-IAI.3B-infected A549 carrier cells in ovarian cancer HEY cells. *P < 0.05; **P < 0.01. (d) Changes in viability of AdE3-IAI.3B-infected A549 carrier cells by infection time. *P < 0.05. (e) Changes of PFU activity of AdE3-IAI.3B-infected A549 carrier cells by infection time. *P < 0.05. (f) Changes of viability of AdE3-IAI.3B-infected A549 carrier cells following liquid nitrogen storage for 3 months. (g) Changes of PFU activity in cells and supernatants of AdE3-IAI.3B-infected A549 carrier cells following liquid nitrogen storage for 3 months.

Mentions: To formulate A549 carrier cells as cancer gene therapy, drug, radiation, infection, and storage conditions were determined. First, to prevent tumorigenicity of A549 carrier cells in patients, radiation dosage was determined. To evaluate the effect of radiation in vitro, A549 cells infected with or without AdE3-IAI.3B were irradiated at 200 Gy. A549 cells infected with AdE3-IAI.3B at 200 multiplicity of infection (MOI) and irradiated A549 cells infected with or without AdE3-IAI.3B at 200 MOI died within 15 days (Figure 2a). To evaluate the effect of radiation in vivo, A549 cells were irradiated and subcutaneously inoculated into nude mice, since A549 cells must be prevented from engrafting into the human body because of the possibility that some of A549 carrier cells might not die without infection with oncolytic adenovirus. Irradiation at levels greater than 200 Gy completely suppressed the tumorigenicity of A549 cells in nude mice (Figure 2b).


Biosafety studies of carrier cells infected with a replication-competent adenovirus introduced by IAI.3B promoter.

Hamada K, Shirakawa T, Terao S, Gotoh A, Tani K, Huang W - Mol Ther Methods Clin Dev (2014)

Infection and storage condition of AdE3-IAI.3B-infected A549 carrier cells. (a) In vitro growth inhibitory effect of A549 carrier cells infected with AdE3-IAI.3B at 200 MOI and irradiated at 200 Gy. (b) Effects of radiation exposure on tumorigenicity of A549 cells subcutaneously transplanted into nude mice (n = 5 per group). (c) Effects of infection time, antiadenovirus antibodies and freeze-thawing on the cytotoxicity of AdE3-IAI.3B-infected A549 carrier cells in ovarian cancer HEY cells. *P < 0.05; **P < 0.01. (d) Changes in viability of AdE3-IAI.3B-infected A549 carrier cells by infection time. *P < 0.05. (e) Changes of PFU activity of AdE3-IAI.3B-infected A549 carrier cells by infection time. *P < 0.05. (f) Changes of viability of AdE3-IAI.3B-infected A549 carrier cells following liquid nitrogen storage for 3 months. (g) Changes of PFU activity in cells and supernatants of AdE3-IAI.3B-infected A549 carrier cells following liquid nitrogen storage for 3 months.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4362384&req=5

fig2: Infection and storage condition of AdE3-IAI.3B-infected A549 carrier cells. (a) In vitro growth inhibitory effect of A549 carrier cells infected with AdE3-IAI.3B at 200 MOI and irradiated at 200 Gy. (b) Effects of radiation exposure on tumorigenicity of A549 cells subcutaneously transplanted into nude mice (n = 5 per group). (c) Effects of infection time, antiadenovirus antibodies and freeze-thawing on the cytotoxicity of AdE3-IAI.3B-infected A549 carrier cells in ovarian cancer HEY cells. *P < 0.05; **P < 0.01. (d) Changes in viability of AdE3-IAI.3B-infected A549 carrier cells by infection time. *P < 0.05. (e) Changes of PFU activity of AdE3-IAI.3B-infected A549 carrier cells by infection time. *P < 0.05. (f) Changes of viability of AdE3-IAI.3B-infected A549 carrier cells following liquid nitrogen storage for 3 months. (g) Changes of PFU activity in cells and supernatants of AdE3-IAI.3B-infected A549 carrier cells following liquid nitrogen storage for 3 months.
Mentions: To formulate A549 carrier cells as cancer gene therapy, drug, radiation, infection, and storage conditions were determined. First, to prevent tumorigenicity of A549 carrier cells in patients, radiation dosage was determined. To evaluate the effect of radiation in vitro, A549 cells infected with or without AdE3-IAI.3B were irradiated at 200 Gy. A549 cells infected with AdE3-IAI.3B at 200 multiplicity of infection (MOI) and irradiated A549 cells infected with or without AdE3-IAI.3B at 200 MOI died within 15 days (Figure 2a). To evaluate the effect of radiation in vivo, A549 cells were irradiated and subcutaneously inoculated into nude mice, since A549 cells must be prevented from engrafting into the human body because of the possibility that some of A549 carrier cells might not die without infection with oncolytic adenovirus. Irradiation at levels greater than 200 Gy completely suppressed the tumorigenicity of A549 cells in nude mice (Figure 2b).

Bottom Line: The use of carrier cells infected with oncolytic viruses in cancer gene therapy is an attractive method because it can overcome viral immunogenicity and induce tumor immunity and significant antitumor activity.Freeze-thawing increased the antitumor effect of A549 carrier cells by promoting the translocation of oncolytic adenovirus particles from the nucleus to the cytoplasm following the rupture of the nuclear membranes.In chronic toxicity tests in rabbits, there were no serious side effects after eight doses of 1.25 × 10(7) cells/kg or less for 4 weeks; a significant immune response is known to elicit increased numbers of antiadenovirus antibodies and enlarge the spleen.

View Article: PubMed Central - PubMed

Affiliation: Department of Obstetrics and Gynecology, School of Medicine, Ehime University , Shitsukawa, Toon, Ehime Japan.

ABSTRACT
The use of carrier cells infected with oncolytic viruses in cancer gene therapy is an attractive method because it can overcome viral immunogenicity and induce tumor immunity and significant antitumor activity. To enable human clinical trials of this treatment, acute and chronic toxicity tests must first be performed to ensure safety. IAI.3B promoter, oncolytic adenovirus AdE3-IAI.3B introduced by IAI.3B promoter, and A549 carrier cells infected with AdE3-IAI.3B were highly active in cancer cells but not in normal cells. Freeze-thawing increased the antitumor effect of A549 carrier cells by promoting the translocation of oncolytic adenovirus particles from the nucleus to the cytoplasm following the rupture of the nuclear membranes. No deaths or abnormal blood test data resulted from acute toxicity tests conducted in nude mice after a single dose. In chronic toxicity tests in rabbits, there were no serious side effects after eight doses of 1.25 × 10(7) cells/kg or less for 4 weeks; a significant immune response is known to elicit increased numbers of antiadenovirus antibodies and enlarge the spleen. From these results, it could be concluded that cancer gene therapy of recurrent solid tumors using carrier cells can be safely trialed in humans.

No MeSH data available.


Related in: MedlinePlus